Single Fluorescent Probe Separately and Continuously Visualize H2S and HClO in Lysosomes with Different Fluorescence Signals.

A complicated relationship between the active small molecules exists in cells. On the organelle level, active small molecules also play an important role in the maintenance of organelle functions and roles. To investigate the relationship of biomolecules in subcellular, it is necessary and critical to develop molecular tools that can track two kinds of associated biomolecules within organelles with multiple fluorescence signals. However, this is still an unmet challenge up to date. Herein, we present the first single-fluorescent probe (Lyso-HA-HS) that can detect oxidative (HOCl) and reductive (H2S) substances within an organelle (lysosomes) with multiresponse signals. The reactions of the new probe with H2S and HOCl simultaneously result in the blue and red channels emissions, respectively, providing different signal responses to the oxidative and reductive substances in the cellular lysosomes. Using a single fluorescent probe, we first achieved dual-channel imaging of the endogenous hypochlorous acid and hydrogen sulfide, respectively, in the lysosomes in the living cells. Moreover, the highly desirable attributes of the probe Lyso-HA-HS (such as high selectivity, good membrane-permeability, and lysosome enrichment ability) may enable it to be used in revealing the relationship of HOCl and H2S in lysosomes.

Single Fluorescent Probe for Dual-Imaging Viscosity and H2O2 in Mitochondria with Different Fluorescence Signals in Living Cells.

Mitochondria, as essential and interesting organelles within the eukaryotic cells, play key roles in a variety of pathologies, and its abnormalities are closely associated with Alzheimer's disease (AD) and other diseases. Studies have shown that the abnormal of viscosity and concentration of hydrogen peroxide in mitochondria were all associated with AD. Accordingly, the detection of viscosity and hydrogen peroxide in mitochondria has attracted great attention. However, it remains a great challenge to explore a single probe, which can dual-detect the viscosity and H2O2 in mitochondria. Herein, in two ways to prevent the twisted internal charge transfer (TICT) process, we designed and sythesized the first dual-detection fluorescent probe Mito-VH that can visualize viscosity and H2O2 in mitochondria with different fluorescence signals in living cells.

A novel collagen/platelet-rich plasma (COL/PRP) scaffold: preparation and growth factor release analysis.

Platelet-rich plasma (PRP) has been widely used in clinical practice for more than 20 years because it causes the release of many growth factors. However, the burst release pattern and short release period of PRP have become obstacles to its application. An optimal controllable release system is an urgent need for researchers. This study investigated whether collagen/PRP (COL/PRP) scaffolds can serve as a vehicle for the controllable release of growth factors. We fabricated a novel scaffold that integrates PRP activated by thrombin or collagen into type I collagen. The mechanical properties, cytotoxicity, and transforming growth factor ß1 (TGF-ß1), platelet derived growth factor (PDGF), fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) content were evaluated. Our results demonstrate that the COL/PRP scaffolds were not cytotoxic to L-929 fibroblasts. The PDGF and FGF content in the thrombin group was at a higher level and lasted for a long period of time. Collagen and thrombin played the same role in the release of TGF-ß1 and VEGF. These data suggest that the novel COL/PRP scaffolds provide a carrier for the controllable release of growth factors and may be used in tissue- regenerative therapies.

A biocompatible chitosan-based fluorescent polymer for efficient H2O2 detection in living cells and water samples.

As a biomarker of oxidative stress, hydrogen peroxide (H2O2) plays a complex role in organisms, including regulating cell signaling, respiration, the immune system, and other life processes. Therefore, it is important to develop a tool that can simply and effectively monitor H2O2 levels in organisms and the environment. In this work, naphthalene fluorophores with a borate structure were introduced into chitosan (CTS) azide, and a CTS-based fluorescence sensor (CTS-HP) was designed for sensitive H2O2 detection. The biocompatibility and degradability of CTS endowed CTS-HP with reduced biotoxicity compared with organic fluorescent dyes, and the substitution degree of fluorophores on the CTS chains was 0.703. The randomly coiled chain structure of the CTS-HP probe enabled the boronic acid recognition sites on the fluorophores to achieve the enrichment of analyte H2O2 through a synergistic effect. Therefore, the probe CTS-HP (10 µg mL-1) exhibited a 21-fold fluorescence enhancement and good detection limit (LOD = 8.98 nM) in H2O2 solution, reaching the maximum fluorescence response faster (within 16 min). The probe also successfully achieved the fluorescence imaging of endogenous and exogenous H2O2 in zebrafish and living cells and labeled the recovery experiment of H2O2 in real water samples (recoveries rates of 90.93-102.9 % and RSD < 3.09 %).

Strategies for measuring concentrations and forms of amyloid-ß peptides.

Alzheimer's disease (AD) is affecting more and more people worldwide without the effective treatment, while the existed pathological mechanism has been confirmed barely useful in the treatment. Amyloid-ß peptide (Aß), a main component of senile plaque, is regarded as the most promising target in AD treatment. Aß clearance from AD brain seems to be a reliably therapeutic strategy, as the two exited drugs, GV-971 and aducanumab, are both developed based on it. However, doubt still exists. To exhaustive expound on the pathological mechanism of Aß, rigorous analyses on the concentrations and aggregation forms are essential. Thus, it is attracting broad attention these years. However, most of the sensors have not been used in pathological studies, as the lack of the bridge between analytical chemist and pathologists. In this review, we made a brief introduce on Aß-related pathological mechanism included in ß-amyloid hypothesis to elucidate the detection conditions of sensor methods. Furthermore, a summary of the sensor methods was made, which were based on Aß concentrations and form detections that have been developed in the past 10 years. As the greatest number of the sensors were built on fluorescent spectroscopy, electrochemistry, and Roman spectroscopy, detailed elucidation on them was made. Notably, the aggregation process is another important factor in revealing the progress of AD and developing the treatment methods, so the sensors on monitoring Aß aggregation processes were also summarized.

A fluorescent probe for detecting H2O2 and delivering H2S in lysosomes and its application in maintaining the redox environments.

Hydrogen peroxide (H2O2) is a reactive oxygen species (ROS) that can be used as a marker for the occurrence of oxidative stress in the organism. Lysosomes serve as intracellular digestive sites, and when the concentration of H2O2 in them is abnormal, lysosomal function is often impaired, leading to the development of diseases. Hydrogen sulfide (H2S) acts as a gaseous signaling molecule that scavenges H2O2 from cells and tissues, thereby maintaining the redox environment of the body. However, most of the reported hydrogen peroxide fluorescent probes so far can only detect H2O2, but cannot maintain the intracellular redox environment. In this paper, an H2O2 fluorescent probe LN-HOD with lysosomal targeting properties was designed and synthesized by combining the H2O2 recognition site with a naphthylamine fluorophore via a thiocarbamate moiety. The probe has the advantages of large Stokes shift (110 nm), high sensitivity and good H2S release capability. The probe LN-HOD can be used to detect H2O2 in cells, zebrafish and plant roots. In addition, LN-HOD detects changes in the concentration of H2O2 in plant roots when Arabidopsis is stressed by cadmium ion (Cd2+). And through its ability to release H2S, it can help to remove excess H2O2 and maintain the redox environment in cells, zebrafish and plant roots. The present work provides new ideas for the detection and assisted removal of H2O2, which contributes to the in-depth study of the cellular microenvironment in organisms.

Near-infrared dual-response fluorescent probe for detection of N2H4 and intracellular viscosity changes in biological samples and various water samples.

N2H4 is a common raw material used in the production of pesticides and has good water solubility, so it may contaminate water sources and eventually enter living organisms, causing serious health problems. Viscosity is an important indicator of the cellular microenvironment and an early warning signal for many diseases. The high reactivity of hydrazine depletes glutathione (GSH) in hepatocytes, causing oxidative stress ultimately leading to significant changes in intracellular viscosity and even death. Therefore, it is particularly important to develop an effective method to detect N2H4 and viscosity in environmental and biological systems. On this basis, we developed two fluorescent probes, BDD and BHD, based on xanthene and 2-benzothiazole acetonitrile. The experimental results show that BHD and BDD have good imaging capabilities for N2H4 in cells, zebrafish and Arabidopsis. BHD and BDD also showed sensitive detection and fluorescence enhancement in the near-infrared region when the intracellular viscosity was changed. Notably, the probe BDD has also successfully imaged N2H4 in a variety of real water samples.

A water-soluble and biocompatible chitosan-based fluorescent probe for real-time monitoring formaldehyde in living cells and zebrafish.

Formaldehyde (HCHO) is a common environmental toxicant that can harm the human respiratory tract and nervous system when exposed for long period of time. As a carcinogen, HCHO also increases the risk of cancer in humans. HCHO can be produced endogenously in living systems and plays an essential role in physiological and biochemical reactions and pathogenesis. Therefore, monitoring the level of HCHO in vivo and in vitro has become the focus of attention. The designed naphthalene fluorophore was introduced onto modified chitosan to prepare a chitosan-based fluorescent probe (CS-FA) for HCHO detection. Compared to other small-molecule probe analogs for the detection of HCHO, the randomly coiled polymer chain of chitosan enabled CS-FA to "enrich" HCHO using the synergistic binding of hydrazino-naphthalimide recognition sites. Thus, the reaction of the analyte with the recognition site was accelerated, resulting in a faster equilibrium fluorescence response (2-3 min) and high sensitivity. In addition, the introduction of biomass material chitosan also improved the biocompatibility of the probe. Then a series of composite materials (test strips and hydrogel) were prepared based on the probe to expand the application form of the probe.

Development of a two-photon fluorescent probe for imaging hydrogen sulfide (H2S) in living cells and zebrafish.

We present a new two-photon fluorescent probe (T-HS) for the detection of H2S. With the addition of hydrogen sulfide, the absorption and fluorescence spectra of the probe show regular changes. The probe exhibited favorable properties, such as large turn-on fluorescence signal, good selectivity and low cytotoxicity. Moreover, the probe T-HS was successfully used for the fluorescence imaging of H2S in live cells and zebrafish.

A turn-on fluorescent probe for sensing N2H4 in living cells, zebrafishes and plant root with a large turn-on fluorescence signal.

Hydrazine (N2H4) plays an important role in industrial production, but it is highly toxic, leaking or exposing it will pollute the environment and cause serious harm to human beings. Therefore, it is necessary to use a simple and effective method to detect N2H4 in environmental systems and organisms. Herein, a novel water-soluble fluorescent probe based on coumarin fluorophore, 2-(7-(diethylamino)-2-oxo-2H-chromen-3-yl)isoindoline-1,3-dione (C-Z1), is reported. The fluorescence intensity of the probe at 530 nm was enhanced gradually with the addition of N2H4, and the maximum enhancement was about 28 times. The probe has good selectivity and sensitivity, the detection limit of hydrazine hydrate is 1.48 × 10-7 M, and the response mechanism of the probe is proved by theoretical calculation and experiment. C-Z1 has been shown to detect N2H4 in a variety of environmental samples, including water, soil, air, cells, zebrafish and plants. In addition, C-Z1 can be made into test strips for easy portability and used for rapid quantitative detection of N2H4 in the field by its distinct change in fluorescence color. Thus, C-Z1 has great potential for the analysis and detection of environmental contaminants.

A highly sensitive and low toxicity cellulose-based fluorescent polymer for H2S detection in cells, zebrafish and food samples.

A cellulose based polymer probe (HC-HS) was prepared for the detection of H2S. HC-HS can be applied to fluorescence imaging of H2S in living cells and zebrafish, and HC-HS was made into test strips to detect H2S produced in the process of food corruption.

Construction of a water-soluble fluorescent probe for copper (II) ion detection in live cells and food products.

The quality of wine can be affected by excess Cu2+ due to the occurrence of oxidation reactions or precipitation. Therefore, it is essential to use simple and effective testing methods to ensure the Cu2+ content in wine. In this work, we designed and synthesized a rhodamine polymer fluorescent probe (PEG-R). The water solubility of PEG-R was improved by the introduction of polyethylene glycol, which improved the performance and broadened its application in the food field. The PEG-R was characterized by high sensitivity, selectivity and fast response to Cu2+ and was able to complete the response process within 30 s, with approximately 29-fold fluorescence enhancement of the probe after exposure to Cu2+, the limit of detection (LOD) was 1.295 × 10-6 M. The probe can be used for the determination of Cu2+ in living cells, zebrafish, white wine and food products, and it was made into practical gels and test strips.

Thermodynamics and conformations in the formation of excited states and their interconversions for twisted donor-substituted tridurylboranes.

We synthesized a series of donor-substituted tridurylboranes containing different types and number of chromophores including 1-pyrene (PB1-3), 3-carbazole (CBC1-3), or substituted p-carbazol-N-phenyl (CBN3a-c) as various donor-acceptor (D-A) molecules. The photophysical and electrochemical properties of these twisted D-A molecules were investigated by means of UV/Vis absorption and fluorescence spectroscopy as well as cyclic voltammetry (CV). Solvent polarity, viscosity, and temperature effects on the fluorescence emission reveal the existence of three types of excited states, and their equilibria and interconversions between three excited states. In increasing order of the charge-separated extent and the conformational change, three excited states are the locally excited (LE) state, the more planar intramolecular charge-transfer (ICT) state, and the more twisted ICT (TICT) state as compared to the ground state. The TICT state undergoes a conformational change with a higher energy barrier over the ICT state. The solvent polarity effect on the state conversion is opposite to the viscosity effect, and temperature effects derive from its resulting changes of polarity and viscosity. For example, the increase of the polarity of the solvent results in excited-state conversions from the LE state to the ICT state, and/or from the ICT to the TICT state, and an increased viscosity leads to the opposite conversions. On the basis of electrochemical and spectral data, thermodynamics of a possible ICT process were estimated, and correlated with the excited-state character. Finally, three excited states have been characterized by the conformation, the photophysical properties, and the thermodynamics of the ICT processes.

Recent Studies on the Preparation and Application of Ionic Amphiphilic Lignin: A Comprehensive Review.

As the second most abundant natural polymer after cellulose, lignin has received considerable attention recently due to its reproducibility, safety, and biodegradability. Studies are now focusing on the development of new lignin applications to replace petroleum-based chemicals. Unfortunately, lignin has several inherent problems, such as poor water solubility and a tendency to agglomerate. However, after chemical modification, lignin can gain new functions through the introduction of new functional groups. For example, amphiphilic lignin is a polymer that is soluble in both water and organic solvents. Amphiphilic lignin polymers can be divided into anionic, cationic, and anionic-cationic amphoteric lignin-based polymers, according to the ions contained in their molecular structure. Amphiphilic lignin polymers also have a wide range of applications in various industrial fields and can be used as wetting agents, detergents, controlled release fertilizers, adsorbents, and emulsifiers. Thus, this article reviews research progress on the synthesis and applications of amphiphilic lignin-derived polymers over the past 10 years, providing a theoretical reference for the utilization of high-added-value and high-performance lignin.

Construction of a unique two-photon fluorescent probe and the application for endogenous CO detection in live organisms.

Carbon monoxide (CO) is one of the most significant signal molecules and plays an important role in regulating human physiological and pathological processes. In this study, a novel Pd-based complex (Pd-BNP-OH) was developed for endogenous CO detection. The structure and morphology of Pd-BNP-OH was characterized by SEM, XPS, and NMR analyses. When Pd-BNP-OH was reacted with CO, a strong fluorescence enhancement at 510 nm was observed. In addition, Pd-BNP-OH exhibited high stability and selectivity toward CO in PBS buffer. In biological experiments, Pd-BNP-OH exhibited little cytotoxicity in cellular environment, and a bright fluorescence turn on was observed in the presence of exogenous CO and endogenous generated CO. The probe was then applied for CO detection in live zebrafish by both one-photon and two-photon excitation. Significantly, Pd-BNP-OH has excellent two-photon property, controllable structure and high biocompatibility. These features enable the probe to detect endogenously generated carbon monoxide in live organisms successfully.

Highly efficient and regiospecific photocyclization of 2,2'-diacyl bixanthenylidenes.

In contrast to the reversible photochemistry of the 2,2'-substituted bixanthenylidenes (1a-f), the photocyclization of 2,2'-diacyl bixanthenylidenes (1g-j) reveals an irreversible process where the initial cyclic intermediate C(E) can undergo a rapid [1,11] hydrogen shift to form stable isomer C'(E) in a degassed solution, which cannot revert to the starting compound, so giving a highly efficient and regiospecific photocyclization.

Fluorescent pyrene-centered starburst oligocarbazoles with excellent thermal and electrochemical stabilities.

A series of pyrene-centered starburst oligocarbazoles (1-3) have been synthesized and well characterized. Based on photophysical, thermal and electrochemical studies in solutions and as thin films, all starburst molecules reveal a sky blue emission with a high efficiency (Φ(F) = 0.99-0.81) and excellent thermal and electrochemical stabilities. As OLED materials, these superior properties are helpful to enhance device stability and lifetime.

Development of a novel NIR viscosity fluorescent probe for visualizing the kidneys in diabetic mice.

Viscosity is an important parameter for evaluating cell health, and abnormal viscosity can cause a variety of intracellular organelle function disorders. The mitochondria are a key organelle in cells, and the viscosity of the mitochondria determines the state of the cell. In this work, we report a novel near-infrared fluorescent probe, referred to as NI-VD, that has a large Stokes-shift and a satisfactory response multiple. NI-VD can sensitively detect changes in cell viscosity in cells and tissues, and it can effectively avoid interference from the overlap of excitation and emission light. The fluorescence spectrum shows that NI-VD has maximum emission peaks at 730 nm, and the fluorescence intensity is amplified with an increase in the solution viscosity. The response from pure PBS solution to glycerol changes by 13-fold. After confirmation in a variety of cell and biological models, NI-VD can detect the changes in viscosity in mitochondria. Most importantly, this study is the first to visualize the differences between the kidneys of diabetic mice and normal mice. This approach is a new solution for the diagnosis and treatment of diabetic nephropathy.

Construction of a dual-response fluorescent probe for copper (II) ions and hydrogen sulfide (H2S) detection in cells and its application in exploring the increased copper-dependent cytotoxicity in present of H2S.

Multiple types of metal ions and active small molecules (reactive nitrogen species, reactive oxygen species, reactive sulfur species, etc.) exist in living organisms. They have connections to each other and can interact and/or interfere with each other. To investigate the relationship of metal ions and active small molecules in living cells, it is necessary and critical to develop molecular tools that can track two kinds of associated certain metal ions and reactive molecules with multiple fluorescence signals. However, this is a challenging task that requires an ingenious molecular design to achieve this goal. Here, we present a fluorescent probe (D-CN) that can offer fluorescence imaging of H2S and copper (II) ions with different response signals. Recognition of H2S and Cu (II) by the new probe can result in green and red emissions, respectively, providing different signal responses to the two substances in living cells and zebrafish. In addition, we used this probe to visually prove that the cytotoxicity of copper ions in living cells increases in the presence of hydrogen sulfide and could lead to cell apoptosis.

A fluorescent probe for specific detection of ß-galactosidase in living cells and tissues based on ESIPT mechanism.

ß-galactosidase is of great significance to living organisms, which is an important marker of primary ovarian cancer and cellular senescence. To detect the activity of ß-galactosidase, a novel fluorescent probe ESIPT-GAL which based on excited state intramolecular proton transfer (ESIPT) mechanism for detecting ß-galactosidase is developed in this work with low background fluorescence and high sensitivity (ΦF = 0.0045-0.2409). The fluorescence intensity at 552 nm of this probe increased by ~ 55 times with ß-galactosidase addition (0-4 U/mL), and its detection limit is very low (3.9 × 10-5 U/mL). In addition, the spectral data (pseudo-first-order rate: 1.303 min-1) and enzyme kinetic parameter (Vmax = 69.5 µΜ•S-1) both show that the probe can achieve rapid response to ß-galactosidase. Moreover, the probe has good water solubility, which ensures that it has good biocompatibility and can be easily applied to detect ß-galactosidase in living cells and tissues. Importantly, the probe ESIPT-GAL can monitor ß-galactosidase in deep mouse tissue sections (90 µm).